title: A reliable amplification technique for the characterization of genomic DNA sequences flanking insertion sequences.
abstract: A simple and efficient ligation-mediated PCR (LMPCR) is described for amplifying DNA adjacent to known sequences. The method uses one primer specific for the known sequence and a second specific for a synthetic linker ligated to restricted genomic DNA. Perkin-Elmer AmpliTaq Gold polymerase is used to minimize non-specific primer annealing and amplification. This LMPCR method was successfully applied to isolate DNA sequences flanking mobile elements present in mycobacterial mutants generated by transposon mutagenesis.
authors.affiliation: Centre national de référence des mycobactéries, Institut Pasteur, Paris, France.
authors.forename: G
authors.initials: G
authors.lastname: Prod'hom
authors.forename: B
authors.initials: B
authors.lastname: Lagier
authors.forename: V
authors.initials: V
authors.lastname: Pelicic
authors.forename: A J
authors.initials: AJ
authors.lastname: Hance
authors.forename: B
authors.initials: B
authors.lastname: Gicquel
authors.forename: C
authors.initials: C
authors.lastname: Guilhot
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers; D004251:DNA Transposable Elements / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis*; D016680:Genome, Bacterial; D016254:Mutagenesis, Insertional / Q000379:methods*; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D016133:Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D004251:DNA Transposable Elements; D004269:DNA, Bacterial